Expanding the Diversity of Unnatural Cell‐Surface Sialic Acids

Luchansky, Sarah J.; Goon, Scarlett; Bertozzi, Carolyn R.

doi:10.1002/cbic.200300789

Cited by 135 publications

(129 citation statements)

References 15 publications

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“…In recent years, small bioorthogonal functional groups, most notably azides and ketones, have been exploited to tag proteins (24)(25)(26)(27), glycans (28)(29)(30)(31), and lipids (32) in cells. These groups are incorporated by the cell's own biosynthetic machinery; the subsequent ligation with reactive probes allows their detection (for a recent review, see ref.…”

mentioning

confidence: 99%

Selective identification of newly synthesized proteins in mammalian cells using bioorthogonal noncanonical amino acid tagging (BONCAT)

Dieterich

Link

Graumann

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

772

756

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In both normal and pathological states, cells respond rapidly to environmental cues by synthesizing new proteins. The selective identification of a newly synthesized proteome has been hindered by the basic fact that all proteins, new and old, share the same pool of amino acids and thus are chemically indistinguishable. We describe here a technology, based on the cotranslational introduction of azide groups into proteins and the chemoselective tagging of azide-labeled proteins with an alkyne affinity tag, to separate and identify, specifically, the newly synthesized proteins in mammalian cells. Incorporation of the azide-bearing amino acid azidohomoalanine is unbiased, not toxic, and does not increase protein degradation. As a first demonstration of the method, we report the selective purification and identification of 195 metabolically labeled proteins with multidimensional liquid chromatography in-line with tandem MS. Furthermore, in combination with leucine-based mass tagging, candidates were immediately validated as newly synthesized proteins. The identified proteins, synthesized in a 2-h window, possess a broad range of biochemical properties and span most functional gene ontology categories. This technology makes it possible to address the temporal and spatial characteristics of newly synthesized proteomes in any cell type.chemical reporter ͉ protein synthesis ͉ proteomics

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mentioning

confidence: 99%

Selective identification of newly synthesized proteins in mammalian cells using bioorthogonal noncanonical amino acid tagging (BONCAT)

Dieterich

Link

Graumann

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

772

756

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“…A series of developments in reporter reactions has made great progress in the imaging of carbohydrates, such as, the azide can be detected by reacting with phosphines via the staudinger ligation, 84 1,3-dipolar cycloaddition between the azide and linear alkynes under catalysis of Cu (abbreviated CuAAC), a variety of interactions of cyclooctynes and azides via Cu-free click chemistry, the reaction of pyradazine or dihydropyrdazine synthesized from tetrazines with various unsaturated compounds via inverse-demand Diels-Alder reaction, and the isonitrile based click chemistry. So far, many carbohydrates have been labeled with bio-orthogonal chemical reporters, such as, sialic acid (Sia) was metabolically labeled with its biosynthetic precursor N-azudoacetylmannisamine (ManNAz), 84,85 N-azudoacetylneuraminic acid (SiaNAz), 85,86 and alkynyl ManNAc. 87 The core GalNAc was labeled by feeding cells or animals with per-O-acetylated N-azidoacetylgalactosamine (Ac 4 GalNAz).…”

Section: Bio-orthogonal Metabolic Labeling For Carbohydratesmentioning

confidence: 99%

Super-resolution imaging in glycoscience: New developments and challenges

Chen

Tong

Wang

2016

J. Innov. Opt. Health Sci.

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Carbohydrates on cell surfaces play a crucial role in a wide variety of biological processes, including cell adhesion, recognition and signaling, viral and bacterial infection, in°ammation and metastasis. However, owing to the large diversity and complexity of carbohydrate structure and nongenetically synthesis, glycoscience is the least understood¯eld compared with genomics and proteomics. Although the structures and functions of carbohydrates have been investigated by various conventional analysis methods, the distribution and role of carbohydrates in cell membranes remain elusive. This review focuses on the developments and challenges of super-resolution imaging in glycoscience through introduction of imaging principle and the available°uorescent probes for super-resolution imaging, the labeling strategies of carbohydrates, and the recent applications of super-resolution imaging in glycoscience, which will promote the super-resolution imaging technology as a promising tool to provide new insights into the study of glycoscience.

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“…4 " 8 NeuAc analogs modified with bio-orthogonal groups are also readily taken up by living cells and incorporated into glycoproteins, allowing subsequent selective chemical manipulations on cell surface for structure-function studies of these glycoproteins. 9 " 11 Recently, sialic acid substituted with a 4-biphenylcarbonyl (BPC) group at C-9 was demonstrated to be a potent inhibitor of CD22, providing renewed interest in exploiting 9-substituted NeuAc analogs as probes of the biology of sialic acid binding proteins. 7,8 CD22 is a negative regulator of B cell activation, mediated by recruitment of SIC domain-containing phosphatase 1 (SHP-1) to the B cell receptor (BCR) through the cytoplasmic domain of CD22.…”

Section: Introductionmentioning

confidence: 99%

Synthesis of 9-Substituted Sialic Acids as Probes for CD22-Ligand Interactions on B Cells

Han

Collins

Paulson

2007

ACS Symposium Series

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As a member of the sialic acid binding immunoglobulin-like lectins (siglecs) family, the B cell protein CD22 binds to NeuAca2-6Gal terminated glycans of glycoproteins on the same cell (in cis) and on adjacent cells (in trans). As a route to develop ligands with altered biological properties, sialic acid analogs with C-9 substitutions were efficiently synthesized from 9-azido-NeuAc, which was obtained in two steps in 65% overall yield from neuraminic acid. 9-Substituted sialic acid analogs were incorporated into cell surface glycoproteins of B cells via the normal cellular biosynthetic pathway, providing unique approaches for the study of CD22-ligand interaction, including in situ photoaffinity crossliking to cis ligands, and modulating the binding affinity with both cis and trans ligands in the cellular context. 2

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Expanding the Diversity of Unnatural Cell‐Surface Sialic Acids

Abstract: Introduction of sialic acid analogues to cells, rather than their upstream N‐acetylmannosamine (ManNAc) precursors, improves the delivery of bioorthogonal functional groups to the cell surface and enables the incorporation of photoactivatable azides into glycoconjugates (see scheme).

Cited by 135 publications

References 15 publications

Selective identification of newly synthesized proteins in mammalian cells using bioorthogonal noncanonical amino acid tagging (BONCAT)

Selective identification of newly synthesized proteins in mammalian cells using bioorthogonal noncanonical amino acid tagging (BONCAT)

Super-resolution imaging in glycoscience: New developments and challenges

Synthesis of 9-Substituted Sialic Acids as Probes for CD22-Ligand Interactions on B Cells

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